Proto-oncogenic isoform A2 of eukaryotic translation elongation factor eEF1 is a target of miR-663 and miR-744
A Vislovukh, G Kratassiouk, E Porto… - British journal of …, 2013 - nature.com
A Vislovukh, G Kratassiouk, E Porto, N Gralievska, C Beldiman, G Pinna, A El'skaya…
British journal of cancer, 2013•nature.comBackground: Eukaryotic translation elongation factor 1A2 (eEF1A2) is a known proto-
oncogene. We proposed that stimulation of the eEF1A2 expression in cancer tissues is
caused by the loss of miRNA-mediated control. Methods: Impact of miRNAs on eEF1A2 at
the mRNA and protein levels was examined by qPCR and western blot, respectively. Dual-
luciferase assay was applied to examine the influence of miRNAs on 3′-UTR of EEF1A2.
To detect miRNA-binding sites, mutations into the 3′-UTR of EEF1A2 mRNA were …
oncogene. We proposed that stimulation of the eEF1A2 expression in cancer tissues is
caused by the loss of miRNA-mediated control. Methods: Impact of miRNAs on eEF1A2 at
the mRNA and protein levels was examined by qPCR and western blot, respectively. Dual-
luciferase assay was applied to examine the influence of miRNAs on 3′-UTR of EEF1A2.
To detect miRNA-binding sites, mutations into the 3′-UTR of EEF1A2 mRNA were …
Abstract
Background:
Eukaryotic translation elongation factor 1A2 (eEF1A2) is a known proto-oncogene. We proposed that stimulation of the eEF1A2 expression in cancer tissues is caused by the loss of miRNA-mediated control.
Methods:
Impact of miRNAs on eEF1A2 at the mRNA and protein levels was examined by qPCR and western blot, respectively. Dual-luciferase assay was applied to examine the influence of miRNAs on 3′-UTR of EEF1A2. To detect miRNA-binding sites, mutations into the 3′-UTR of EEF1A2 mRNA were introduced by the overlap extension PCR.
Results:
miR-663 and miR-744 inhibited the expression of luciferase gene attached to the 3′-UTR of EEF1A2 up to 20% and 50%, respectively. In MCF7 cells, overexpression of miR-663 and miR-744 reduced the EEF1A2 mRNA level by 30% and 50%. Analogous effects were also observed at the eEF1A2 protein level. In resveratrol-treated MCF7 cells the upregulation of mir-663 and mir-744 was accompanied by downregulation of EEF1A2 mRNA. Both miRNAs were able to inhibit the proliferation of MCF7 cells.
Conclusion:
miR-663 and miR-744 mediate inhibition of the proto-oncogene eEF1A2 expression that results in retardation of the MCF7 cancer cells proliferation. Antitumour effect of resveratrol may include stimulation of the miR-663 and miR-744 expression.
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